COPD is a condition characterised by chronic inflammation in which macrophages arethought to play a central role. Corticosteroids are the most widely used antiinflammatoryagents to treat COPD. There is a subset of inflammatory mediatorswhich are corticosteroid insensitive and so there is a need for novel anti-inflammatorytherapies to treat COPD. One such target is the liver X receptor (LXR), a cholesterolsensing nuclear hormone receptor with anti-inflammatory properties. Before investigating the anti-inflammatory potential of LXR, I aimed to validate thealveolar macrophage in vitro culture model. I investigated the effect of differentculture times on unstimulated and stimulated cytokine release, dexamethasoneinhibition of cytokine release, and transcription factor phosphorylation in alveolarmacrophages from COPD patients and controls. I found that freshly isolatedmacrophages release higher levels of cytokines, are less responsive to dexamethasoneand have increased levels of phosphorylated p38 MAPK.I next investigated LXR gene and protein expression levels in alveolar macrophages andwhole lung tissue from COPD patients and controls, the effect of LXR activation on thesuppression of inflammatory mediators from LPS stimulated COPD alveolarmacrophages, and the effect of LXR activation on the induction of genes associatedwith alternative macrophage polarisation. The levels of LXR mRNA were significantlyincreased in whole lung tissue extracts in COPD patients and smokers compared tonever smokers. The expression of LXR protein was significantly increased in smallairway epithelium and alveolar epithelium in COPD patients compared to controls. Nodifferences in LXR mRNA and protein levels were observed in alveolar macrophagesbetween patient groups. The LXR agonist GW3965 significantly induced the expressionof the LXR dependent genes ABCA1 and ABCG1 in alveolar macrophage cultures. InLPS stimulated alveolar macrophages, GW3965 suppressed the production of CXCL10and CCL5, whilst stimulating IL-10 production. GW3965 did not significantly suppressthe production of TNFα, IL-1β, or CXCL8. The major finding is that LXR activation hasanti-inflammatory effects on CXC10, CCL5 and IL-10 production from alveolarmacrophages. Finally, I investigated whether dysfunctional lipid homeostasis is a feature of COPDalveolar macrophages. I found that alveolar macrophages from current smokers withand without COPD had increased levels of neutral lipids compared to never smokersand ex-smokers with and without COPD. Dysfunctional lipid homeostasis may play arole in COPD.

Neutrophils are key effector cells of the innate immune system. They employ a number of powerful ‘weapons’ to eliminate pathogens, including an array of destructive proteins packaged into distinctive granule subsets. In addition to their microbicidal activity, these granule proteins are capable of causing substantial tissue damage if inappropriately deployed. To mitigate against this possibility, most physiological stimuli induce minimal extracellular degranulation. Sites of inflammation and infection are usually hypoxic, and it has been shown that oxygen depletion compromises neutrophil function by impairing the generation of reactive oxygen species and hence bacterial killing. The key finding reported in this thesis is that hypoxia substantially increases the release of all neutrophil granule subsets, as measured by the release of (active) hallmark proteins (elastase, myeloperoxidase, lactoferrin and matrix metalloproteinase-9). In consequence, supernatants from hypoxic neutrophils induced substantially more damage to lung epithelial cell layers than supernatants from neutrophils cultured under normoxic conditions; this damage was protein- and protease-dependent. This pattern of damage was seen consistently across lung adenocarcinoma-derived epithelial cells, primary immortalised lung epithelial cells, and primary human bronchial epithelial cells grown in physiological air-liquid interface culture. Surprisingly, the mechanism of hypoxia-augmented degranulation was found to be independent of protein synthesis and specifically, of the transcription factor HIF-1α (the ‘master-regulator’ of hypoxic responses); thus, hypoxia did not affect mRNA transcript or protein abundance of the major granule components, and hypoxia mimetics failed to recapitulate the phenotype. Inhibition of the key pathways known to be involved in neutrophil degranulation, including, phosphatidylinositol 3-kinase and phospholipase C, but not calcium flux prevented augmented granule release under hypoxia In conclusion, hypoxia induces a destructive neutrophil phenotype, with increased release of multiple histotoxic proteases. This may contribute to tissue injury and disease pathogenesis in a range of clinically important conditions.

The impact of Chronic Obstructive Pulmonary Disorder is material in the dentistry field which not only centers its efforts on managing the oral health of the afflicted patients, prescribing proper medications, but also on raising awareness mainly through dental hygiene and smoking cessation. This literature review will explore the status and accomplishments of the field of dentistry with regards to battling COPD. The first section of the paper discusses the nature and dynamics of the disease which ranks between the third and fourth most common cause of death in the United States alone. Certain data trends from 1970 until 2002 showed double the death rates caused by the disease, and that exposure to environmental or occupational risk factors (e.g. dust, fumes, etc.) must be seriously considered for the effective management of COPD. The pressure of COPD management is great among dental practitioners who must ensure the safety of patients by providing the most suitable drug regimen and having a well-coordinated dental practice (i.e. with other health practitioners) to avoid exacerbation of the disease which often leads to hospitalization. The latter is often the result of oral devices or materials that are often comprised of small objects which might be swallowed or aspirated into the oropharynx of the patient when proper positioning during dental check-ups is not properly performed. Dentists must also make sure that they are able to gain the patient’s confidence and to avoid patient reluctance during dental care. Numbers and statistics also reveal that an estimated 24 million of the population in the United States is afflicted by COPD, and that is from 2015 alone. It is also prevalent among the male population which has a death rate of 83 per 100,000, while the death rate among women is 57 per 100,000. Immediate treatment of COPD is also emphasized to prevent hospitalization rates from rising, and as far as dental practice is concerned, they are often aware of CODP patients which number approximately 130 out of 2000. Smoking cessation is considered the best preventive measure in COPD management. Educating patients on the real dangers involved with smoking will help encourage them to quit while promoting better oral health. The field of dentistry is constantly responding to the challenges of COPD, and significant breakthroughs have already been achieved due to research efforts on COPD management. Through professional coordination with other fields in the healthcare system, it is with great hope that dental practitioners will contribute more to the alleviation of patient conditions and providing improved quality of life.

Dyspné som uppstår vid Chronic Obstructive Pulmonary Disease, COPD, påverkar patienter fysiskt, psykiskt och socialt på ett negativt sätt. Att som sjuksköterska stödja patienten i copingstrategier där patienten lär sig hantera sin dyspné, anpassa sig till sjukdomen och kompensera för begränsningar kan tänkas hjälpa dem att hantera sjukdomen och dess symtom. Syftet med studien var att identifiera och sammanställa patienters erfarenheter av verksamma copingstrategier vid dyspné vid COPD. I denna beskrivande litteraturstudie som utgår från Fribergs analysprocess, granskas och analyseras tolv kvalitativa artiklar. Analysen resulterar i tre identifierade huvudteman, med tillhörande fem subteman som tillsammans sammanfattar patienters erfarenheter av verksamma copingstrategier vid dyspné vid COPD. Identifierade huvudteman i arbetet är fysiska copingstrategier, psykiska copingstrategier och sociala copingstrategier. I den fysiologiska delen upplevde patienterna andningsövningar och handhållen fläkt avlastande. I den psykiska delen ses ett samband mellan upplevelsen av dyspnén i relation till sjukdoms acceptans. I den sociala delen sågs närstående och vården vara av betydelse såsom beröring av närstående och att ta emot hjälp av närstående och vården i det dagliga livet. En tillförlitlig nåbar vård sågs också som en verksam copingstrategi.

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第12956号 / 論医博第2098号 / 新制||医||1011(附属図書館) / 32355 / 明治鍼灸大学大学院鍼灸学研究科鍼灸学専攻 / (主査)教授 伊達 洋至, 教授 佐藤 俊哉, 教授 森田 智視 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

Chronic obstructive pulmonary disease (COPD) is a leading cause of global morbidity and mortality, with the potential to afflict as many as half of the 1.1 billion smokers in the world. The inflammatory response to cigarette smoke is believed to mediate the progressive and irreversible loss of lung function that characterizes COPD. The greatest burden of the disease arises from episodes of worsened symptoms and inflammation, usually triggered by microbial infection. Currently, the mechanisms that drive cigarette smoke-induced inflammation are being elucidated but ambiguity remains about this response and the inflammatory response engaged in a smoke-exposed lung experiencing a microbial infection. This thesis sought to investigate inflammatory mediators induced by cigarette smoke and those induced by bacteria, the most common cause of infectious exacerbations of COPD, in the context of smoke exposure. In chapter two we investigated the role of Breast Regression Protein-39 (BRP-39), a gene commonly observed to be increased under inflammatory conditions, in the inflammatory response to cigarette smoke. In order to determine the mechanisms of BRP-39 induction, its expression and inflammation was assessed in IL-13, IL-18, and IL-1R1 deficient mice. BRP-39 was found to be redundant in cigarette smoke-induced inflammation, but these data confirmed that IL-1R1 was a crucial mediator of this response. After examining the inflammatory response elicited by smoke alone, we investigated the importance of IL-1 signaling in a model of bacterial exacerbation of cigarette smoke-induced inflammation. We found that the exacerbated neutrophilia that typifies the response of a smoke-exposed lung to bacteria was dependent on IL-1α-mediated production of the CXCR2 ligand CXCL5. This study identified the unique phenomenon that cigarette smoke primes alveolar macrophages to produce excessive amounts of IL-1α in response to bacterial stimuli. The purpose of the final study of the thesis was to more comprehensively characterize the extent to which cigarette smoke changes the phenotype of macrophages. Examining total gene expression by microarray found that smoke-exposed alveolar macrophages were in a proliferative state expressing a unique profile of inflammatory mediators. Further analysis revealed that this was likely the result of a pulmonary environment rich in growth factors. Taken together, these data provide detail to the understanding of the biological process of inflammation that drives the pathogenesis of COPD. These studies identify a phenomenon that predisposes smokers to experience more severe responses to bacteria and reinforces the targeting of IL-1 signaling in the treatment of COPD. / Thesis / Doctor of Philosophy (Medical Science)

No description available.

Introduction: Takotsubo cardiomyopathy or stress cardiomyopathy is a syndrome of transient left ventricular (LV) dysfunction mimicking myocardial infarction, but lacking obstruction of coronary artery disease (CAD) or acute plaque rupture. A characteristic differentiation from CAD is that regional motional abnormality extends beyond a territory perfused with a single epicardial coronary artery. Clinically, it is characterized by apical ballooning of the LV due to due to depression of mid and apical segments, with hyperkinesis of cardiac basal walls. Women are affected more than men, predominantly in the postmenopausal age. Case Report: A 54-year-old Caucasian female with a history of COPD, hypertension, uncontrolled diabetes mellitus, hyperlipidemia, depression and ongoing tobacco use presented with complaints of worsening shortness of breath two days prior to admission. She denied chest pain, worsened cough, palpitations, nausea or vomiting. On examination, she was in distress and anxious, with labored breathing. Upon examining the chest, decreased air entry was present in both lung fields with bibasilar wheezing. Initial lab tests showed mild respiratory acidosis, with pH of 7.24, pCO2 of 47.4 and pO2 of 65. Troponins on the day of admission was Soon after admission, she started complaining of severe right neck pain. Repeat EKG revealed localized lateral J point, anteroseptal q waves and 4mm ST-segment elevation in leads V3 and V4 reciprocal changes and without chest pain. Repeat troponins were slightly elevated to 0.42 ng/ml and CK-MB was elevated to 20.2 ng/ml. A transthoracic echocardiogram showed regional abnormalities in left ventricle with the apex, mid to distal septum and the anterior part of septum was akinetic. Discussion: Takotsubo cardiomyopathy presents in 1 to 2 percent of troponin-positive acute coronary syndrome (ACS) with various clinical manifestations and various outcomes. Some patients have favorable outcomes based on their clinical performance and extent of cardiac muscle involvement. As in the case we presented, this syndrome can be entirely idiopathic, without a definitive underlying cause. Supportive management while hospitalized and early identification of complications improve the prognosis.

High-resolution computed tomography (HRCT) of the chest is commonly used in the diagnosis of a variety of lung diseases. Structural changes associated with clinical characteristics of disease may also define specific disease-associated physiologic states that may provide insights into disease pathophysiology. Gene expression profiling is potentially a useful adjunct to HRCT to identify molecular correlates of the observed structural changes. However, it is difficult to directly access diseased distal airway or lung parenchyma routinely for profiling studies. Previously, we have profiled bronchial airway in normal-appearing epithelial cells at the mainstem bronchus, detecting distinct gene expression alterations related to the clinical diagnosis of chronic obstructive pulmonary disease (COPD) and lung cancer. These gene expression alterations offer insights into the molecular events related to diseased tissue at more distal airways and in the parenchyma, which we hypothesize are due to a field-of-injury effect. Here, we expand this prior work by correlating airway gene expression to COPD and bronchiectasis phenotypes defined by HRCT to better understand the pathophysiology of these diseases. Additionally, we classified pulmonary nodules as malignant or benign by combining HRCT nodule imaging characteristics with gene expression profiling of the nasal airway. First, we collected brushing samples from the main-stem bronchus and assessed gene expression alterations associated with COPD phenotypes defined by K-means clustering of HRCT-based imaging features. We found three imaging clusters, which correlated with incremental severity of COPD: preserved, interstitial predominant, and emphysema predominant. 357 genes were differentially expressed between the normal and the emphysema predominant clusters. Functional analysis of the differentially expressed genes suggests a possible induction of inflammatory processes and repression of T-cell related biologic pathways, in the emphysema predominant cluster. We then discovered gene expression alterations associated with radiographic evidence of bronchiectasis (BE), an underdiagnosed obstructive pulmonary disease with unclear pathophysiology. We found 655 genes were differentially expressed in bronchial epithelium from individuals with radiographic evidence of BE despite none of the study participants having a clinical BE diagnosis. In addition to biological pathways that had been previously associated with BE, novel pathways that may play important roles in BE initiation were also discovered. Furthermore, we leveraged an independent single-cell RNA-sequencing dataset of the bronchial epithelium to explore whether the observed gene expression alterations might be cell-type dependent. We computationally detected an increased presence of ciliated and deuterosomal cells, as well as a decreased presence of basal cells in subjects with widespread radiographic BE, which may reflect a shift in the cellular landscape of the airway during BE initiation. Finally, we identified gene expression alterations within the nasal epithelium associated with the presence of malignant pulmonary nodules. A computational model was constructed for determining whether a nodule is malignant or benign that combines gene expression and imaging features extracted from HRCT. Leveraging data from single-cell RNA sequencing, we found genes increased in patients with lung cancer are expressed at higher levels within a novel cluster of nasal epithelial cells, termed keratinizing epithelial cells. In summary, we leveraged gene expression profiling of the proximal airway and discovered novel biological pathways that potentially drive the structural changes representative of physiologic states defined by chest HRCT in COPD and BE. This approach may also be combined with chest HRCT to detect weak signals related to malignant pulmonary nodules. / 2024-02-03T00:00:00Z

Chronic Obstructive Pulmonary Disease (COPD), which is characterized by limitation of pulmonary air flow, is now the third major cause of death worldwide. Barrero et al. have reported that the elevation of extracellular hyperacetylated histone H3.3 in the lungs of COPD patients is associated with cytotoxicity and disease progression. They found that extracellular hyperacetylated H3.3 was cytotoxic to lung structural cells and resistant to proteasomal degradation, and that mouse antibodies to either the C- or N- termini of H3.3 could partially reverse H3.3 toxicity in vitro. Thus, we hypothesize that human antibodies directed against H3.3 may be effective biotherapeutics useful to control progression of COPD in vivo. The discovery and development of human monoclonal antibodies (mAbs) is a fast growing field of biotherapeutics. In addition to full length mAbs, antibody fragments also have been used in antibody discovery research. We have used phage display technology in this project to discover human anti-H3.3 antibody Fab fragments. This technology utilizes genetically engineered phage particles containing genes encoding diverse Fab fragments displayed on the particles. The “Ylanthia” library from MorphoSys AG, a synthetic fully human Fab antibody phage display library with 1.3 x 1011 independent clones, was panned against purified recombinant human H3.3 immobilized on 96-well plates. Seven H3.3-binding Fab fragments with unique DNA sequences were isolated after four rounds of panning. Following their expression in E.coli and purification, Fab purities and electrophoretic mobilities were evaluated on SDS-PAGE. The concentration-dependent binding activities of all seven Fabs to human H3.3 were tested by ELISA. All seven Fabs were shown by ELISA to bind H3.3 but not histones 2A, 2B or 4. Since H3.3 is localized to the nucleus, western blotting was used to demonstrate that seven Fabs recognize purified, recombinant H3.3 and denatured natural histone(s) from nuclear extracts of human 293T cells. In order to characterize these molecules further, biological activity assays will be done to test their potential to reverse the toxic effects of H3.3 in cell culture. If these Fabs prove active in cell culture, they will be converted to IgGs and tested in animal models as potential biotherapeutics for COPD. / Pharmaceutical Sciences